14‐3‐3 proteins associate with the regulatory phosphorylation site of spinach leaf nitrate reductase in an isoform‐specific manner and reduce dephosphorylation of Ser‐543 by endogenous protein phosphatases

FEBS Letters

Huber, Steven C.¹  Athwal, Gurdeep S.¹  Huber, Joan L.¹  Wu, Ke²  Ferl, Robert J.²  Bachmann, Markus¹ 

[1] US Department of Agriculture, Agricultural Research Service, and Departments of Horticulture (MB, JLH, GSA), Crop Science and Botany (SCH), North Carolina State University, Raleigh, NC 27695-7631, USA;Department of Horticultural Sciences, University of Florida, Gainesville, FL 32611-0690, USA

关键词: Binding site;    14-3-3 Protein;    Inhibitor protein;    Isoform specificity;    Nitrate reductase;    Protein phosphatase;    Regulatory phosphorylation site;    IP;    inhibitor protein;    NR;    NADH;    nitrate reductase;    NRk;    NR;    kinase;    NR-PP;    NR-protein phosphatase;   

DOI  :  10.1016/S0014-5793(96)01188-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Three lines of evidence indicate that the 14-3-3 proteins that inactivate the phosphorylated form of spinach leaf NADH:nitrate reductase (NR) bind to the enzyme at the regulatory phosphorylation site (Ser-543). First, a phosphorylated synthetic peptide based on the regulatory site can prevent and also reverse the inactivation of phospho-NR caused by 14-3-3 proteins. Second, sequence-specific and phosphorylation-dependent binding of the aforementioned synthetic peptide to the 14-3-3 proteins was demonstrated in vitro. Third, 14-3-3 proteins were required for the ATP-dependent phosphorylation of NR (as assessed by activity measurements) in the presence of NR-kinase and leaf protein phosphatases. Lastly, we demonstrate specificity of recombinant Arabidopsis 14-3-3 isoforms in the interaction with phospho-NR: ω > χ > ν ⋙ gf, ψ.

【 授权许可】

Unknown